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Betulinic acid attenuates liver fibrosis by inducing autophagy via the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway

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Abstract

The present study was designed to investigate the effects of betulinic acid on human hepatic stellate cells in vitro and C57BL/6 mice in vivo, as well as the signaling pathways involved. In this study, we explored the effects of betulinic acid on expression of alpha smooth muscle actin and autophagy-related proteins. Betulinic acid reduced pathological damage associated with liver fibrosis, as well as serum platelet-derived growth factor and serum hydroxyproline levels. Furthermore, betulinic acid downregulated the expression of alpha smooth muscle actin and type I collagen in mouse liver and upregulated the expression of microtubule-associated protein light chain 3B and autophagy-related gene 7 at the gene and protein levels. LC3II expression was increased and alpha smooth muscle actin expression was decreased in betulinic acid-treated hepatic stellate cells. Interventions with bafilomycin A1 and mCherry-GFP-LC3 adenoviruses promoted the formation of autophagosomes in hepatic stellate cells and the development of autophagic flow. Our study found that mitogen-activated protein kinase/extracellular signal-regulated kinase may be involved in the effects of betulinic acid on liver fibrosis. The present study suggests that betulinic acid has anti-hepatic fibrosis activity by inducing autophagy and could serve as a promising new agent for treating hepatic fibrosis.

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Abbreviations

BA:

Betulinic acid

PDGF:

Platelet-derived growth factor

3-MA:

3-Methyladenine

BafA1:

Bafilomycin A1

TEM:

Transmission electron microscopy

PBS:

Phosphate-buffered saline

MAPK:

Mitogen-activated protein kinase

P70S6K:

Ribosomal protein S6 kinase

ERK:

Extracellular-regulated protein kinases

Col:

Colchicine

LC3:

Microtubule-associated protein 1 light chain 3

ATG7:

Autophagy-related protein 7

α-SMA:

Alpha smooth muscle actin antibody

COL-1:

Collagen I

PI3K:

Phosphatidylinositol 3-kinase

mTOR:

The mammalian target of rapamycin

ECM:

Extracellular matrix

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81603501 and 81774170), Science and Technology Planning Project of Guangdong Province (2014A020221097), Administration of Traditional Chinese Medicine of Guangdong Province (20162087), Science and Technology Planning Project of Guangzhou City (201508020014 and 201707010080), the Natural Science Foundation of Guangdong Province (2018B030306012 and 2017A030313903), the Scientific Research Initiative Program of Southern Medical University (LX2015N003 and CX2017N001), Combined Science Technology Project of Guangdong Provincial Department of Science and Technology and Guangdong Provincial Academy of Traditional Chinese Medicine (2014A020221011); Guangdong Province Bureau of Traditional Chinese Medicine Scientific Research Project (20161161).

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  1. Yuan Liu and Yanmeng Bi contributed equally to this work and are co-first authors.

Authors and Affiliations

  1. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong, China

    Yuan Liu, Yanmeng Bi, Chan Mo, Ting Zeng, Sha Huang, Lei Gao, Xuegang Sun & Zhiping Lv

  2. The Key Laboratory of Molecular Biology, State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong, China

    Lei Gao & Xuegang Sun

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  1. Yuan Liu
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Contributions

ZL and XS participated in the conception and design of the study; YL and YB participated in generation, collection, assembly, and interpretation of data; CM and TZ participated in drafting and revision of the manuscript; SH and LG participated in statistical analysis; ZL obtained funding.

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Correspondence to Xuegang Sun or Zhiping Lv.

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Liu, Y., Bi, Y., Mo, C. et al. Betulinic acid attenuates liver fibrosis by inducing autophagy via the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway. J Nat Med 73, 179–189 (2019). https://doi.org/10.1007/s11418-018-1262-2

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